Improved assay to detect Plasmodium falciparum using an uninterrupted, semi-nested PCR and quantitative lateral flow analysis.
| Autor: | Ongagna-Yhombi SY; Department of Basic Science, NYU College of Dentistry, New York, NY 10010, USA., Corstjens P, Geva E, Abrams WR, Barber CA, Malamud D, Mharakurwa S |
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| Jazyk: | angličtina |
| Zdroj: | Malaria journal [Malar J] 2013 Feb 22; Vol. 12, pp. 74. Date of Electronic Publication: 2013 Feb 22. |
| DOI: | 10.1186/1475-2875-12-74 |
| Abstrakt: | Background: A rapid, non-invasive, and inexpensive point-of-care (POC) diagnostic for malaria followed by therapeutic intervention would improve the ability to control infection in endemic areas. Methods: A semi-nested PCR amplification protocol is described for quantitative detection of Plasmodium falciparum and is compared to a traditional nested PCR. The approach uses primers that target the P. falciparum dihydrofolate reductase gene. Results: This study demonstrates that it is possible to perform an uninterrupted, asymmetric, semi-nested PCR assay with reduced assay time to detect P. falciparum without compromising the sensitivity and specificity of the assay using saliva as a testing matrix. Conclusions: The development of this PCR allows nucleic acid amplification without the need to transfer amplicon from the first PCR step to a second reaction tube with nested primers, thus reducing both the chance of contamination and the time for analysis to < two hours. Analysis of the PCR amplicon yield was adapted to lateral flow detection using the quantitative up-converting phosphor (UCP) reporter technology. This approach provides a basis for migration of the assay to a POC microfluidic format. In addition the assay was successfully evaluated with oral samples. Oral fluid collection provides a simple non-invasive method to collect clinical samples. |
| Databáze: | MEDLINE |
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